APOL1 - Antibodies - The Human Protein Atlas


	Antibody HPA018885	Antibody CAB056156

ANTIBODY INFORMATION
Provider	Atlas Antibodies Sigma-Aldrich	Atlas Antibodies
Product name	HPA018885	AMAb90530
Host species	Rabbit	Mouse
Clonalityⁱ The antibodies are designated mAB for monoclonal and pAb for polyclonal.	pAb	mAb
Concentration	0.12 mg/ml	Not known
Purity	Affinity purified using the PrEST-antigen as affinity ligand	Protein A/G
Released in versionⁱ The release of the Human Protein Atlas in which the antibody was first published.	4.1	10.0
Referencesⁱ References to publications in which the antibody has been used.	17
Proper citation	Atlas Antibodies Cat#HPA018885, RRID:AB_1844953	Atlas Antibodies Cat#AMAb90530, RRID:AB_2665576
Validation summaryⁱ All assays through which the antibody has been validated. Assays&annotation provide a detailed description of the different assays. The pie-charts indicate degree of validation.	N/A ICC IHC N/A WB PA	N/A ICC IHC WB N/A PA

IMMUNOHISTOCHEMISTRYⁱ Immunohistochemistry is used for validating antibody reliability by assessing staining pattern in 44 normal tissues. Validation scores include Enhanced, Supported, Approved and Uncertain.
Validationⁱ Results of validation by standard or enhanced validation based on assessment of antibody performance in 44 normal tissues. Standard validation results in scores Supported, Approved or Uncertain. An image representative of the antibody staining pattern is shown. Enhanced validation results in the score Enhanced and includes two methods: Orthogonal validation and Independent antibody validation. For orthogonal validation, representative images of high and low expression are shown. For independent antibody validation, four images of each independent antibody are displayed.	Supportedⁱ Immunohistochemistry is used for validating antibody reliability by assessing staining pattern in 44 normal tissues. Validation scores include Enhanced, Supported, Approved and Uncertain. Immunohistochemical staining of human vulva/anal skin shows distinct positivity in plasma. Skin 2	Supportedⁱ Immunohistochemistry is used for validating antibody reliability by assessing staining pattern in 44 normal tissues. Validation scores include Enhanced, Supported, Approved and Uncertain. Immunohistochemical staining of human colon shows distinct positivity in plasma. Colon

Retrievalⁱ Antigen retrieval is a method used to restore/retrieve the epitope (antibody bidning region) of the target protein, cross-linked, and thus masked, during tissue preserving fixative treatment of the tissues.	HIER pH6	HIER pH6
Antibody dilution	1:500	1:3500
Literature conformityⁱ Conformance of the expression pattern with available gene/protein characterization data in scientific literature and data from bioinformatic predictions. UniProt is used as the main source of gene/protein characterization data and when relevant, available publications and other sources of information are researched in depth. Extensive or sufficient gene/protein data requires that there is evidence of existence on a protein level and that a substantial quantity of published experimental data is available from literature and public databases. Limited protein/gene characterization data does not require evidence of existence on a protein level and refers to genes for which only bioinformatic predictions and scarce published experimental data is available.	Consistent with extensive gene/protein characterization data.	Consistent with extensive gene/protein characterization data.
RNA consistencyⁱ Consistency between immunohistochemistry data and consensus RNA levels is divided into five different categories: i) High consistency, ii) Medium consistency, iii) Low consistency, iv) Very low consistency, and v) Cannot be evaluated.	No tissue staining or RNA expression data available for comparison.	No tissue staining or RNA expression data available for comparison.
WESTERN BLOTⁱ A Western blot analysis is performed on a panel of human tissues and cell lines to evaluate antibody specificity. For antibodies with unreliable result a revalidation using an over-expression lysate is performed.
Validationⁱ Western Blot is used for quality control of the polyclonal antibodies generated in the project. After purification, the antibodies are used to detect bands in a setup of lysate and different tissues. The result is then scored Enhanced, Supported, Approved, or Uncertain. Enhanced validation includes five different methods: Genetic validation, Recombinant expression validation, Independent antibody validation, Orthogonal validation and Capture MS validation.	Uncertainⁱ The staining of an antibody is evaluated by Western Blot through analysis of samples from different cell lysates. A supportive score is given if band(s) of predicted size in kDa (+/-20%) is detected. Weak band of predicted size but with additional bands of higher intensity also present. Analysis performed using a standard panel of samples.	Supportedⁱ The staining of an antibody is evaluated by Western Blot through analysis of samples from different cell lysates. A supportive score is given if band(s) of predicted size in kDa (+/-20%) is detected. Band of predicted size in kDa (+/-20%) with additional bands present. Analysis performed using a standard panel of samples. 250 130 100 70 55 35 25 15 10

Antibody dilution	1:250	1:1000
PROTEIN ARRAY
Validationⁱ A protein array containing 384 different antigens including the antibody target is used to analyse antibody specificity. Depending on the array interaction profile the antibody is scored as Supported, Approved, or Uncertain.	Supported Pass with single peak corresponding to interaction only with its own antigen. Antibody specificity analysis with protein arrays. Predicted and matching interactions are shown in green.	N/A

Antibody dilution	1:3000
RELEVANT PUBLICATIONS
	APOL1 localization in normal kidney and nondiabetic kidney disease Madhavan SM et al J Am Soc Nephrol 2011;22(11):2119-28
	APOL1 null alleles from a rural village in India do not correlate with glomerulosclerosis Johnstone DB et al PLoS One 2012;7(12):e51546
	Localization of APOL1 protein and mRNA in the human kidney: nondiseased tissue, primary cells, and immortalized cell lines Ma L et al J Am Soc Nephrol 2015;26(2):339-48 Application: WB
	Exon 4-encoded sequence is a major determinant of cytotoxicity of apolipoprotein L1 Khatua AK et al Am J Physiol Cell Physiol 2015;309(1):C22-37 Application: WB
	Biogenesis and cytotoxicity of APOL1 renal risk variant proteins in hepatocytes and hepatoma cells Cheng D et al J Lipid Res 2015;56(8):1583-93 Application: IP
	BH3 domain-independent apolipoprotein L1 toxicity rescued by BCL2 prosurvival proteins Heneghan JF et al Am J Physiol Cell Physiol 2015;309(5):C332-47 Application: WB
	APOL1-Mediated Cell Injury Involves Disruption of Conserved Trafficking Processes Kruzel-Davila E et al J Am Soc Nephrol 2017;28(4):1117-1130 Application: WB
	Transgenic expression of human APOL1 risk variants in podocytes induces kidney disease in mice Beckerman P et al Nat Med 2017;23(4):429-438 Application: WB
	A tripartite complex of suPAR, APOL1 risk variants and αβ integrin on podocytes mediates chronic kidney disease Hayek SS et al Nat Med 2017;23(8):945-953 Application: IHC
	Intracellular APOL1 Risk Variants Cause Cytotoxicity Accompanied by Energy Depletion Granado D et al J Am Soc Nephrol 2017;28(11):3227-3238 Application: WB
	APOL1 variants change C-terminal conformational dynamics and binding to SNARE protein VAMP8 Madhavan SM et al JCI Insight 2017;2(14): Application: ICC-IF, IHC, WB
	APOLs with low pH dependence can kill all African trypanosomes Fontaine F et al Nat Microbiol 2017;2(11):1500-1506 Application: WB
	Blocking the 5' splice site of exon 4 by a morpholino oligomer triggers APOL1 protein isoform switch Cheatham AM et al Sci Rep 2018;8(1):8739 Application: WB
	APOL1 risk allele RNA contributes to renal toxicity by activating protein kinase R Okamoto K et al Commun Biol 2018;1:188 Application: WB
	A null variant in the apolipoprotein L3 gene is associated with non-diabetic nephropathy Skorecki KL et al Nephrol Dial Transplant 2018;33(2):323-330 Application: IP
	APOL1 renal risk variants promote cholesterol accumulation in tissues and cultured macrophages from APOL1 transgenic mice Ryu JH et al PLoS One 2019;14(4):e0211559 Application: IHC
	Antisense oligonucleotide treatment ameliorates IFN-γ-induced proteinuria in APOL1-transgenic mice Aghajan M et al JCI Insight 2019;4(12): Application: WB
ANTIGEN INFORMATION
Antigen	Recombinant protein fragment	Recombinant protein
Length (aa)	125
Antigen sequence	`SNFLSLAGNTYQLTRGIGKDIRALRRARANLQSVPHASASRPRVTEPISA ESGEQVERVNEPSILEMSRGVKLTDVAPVSFFLVLDVVYLVYESKHLHEG AKSETAEELKKVAQELEEKLNILNN`
Matching transcripts	APOL1-201 - ENSP00000317674 [100%] APOL1-202 - ENSP00000380448 [100%] APOL1-204 - ENSP00000388477 [100%] APOL1-205 - ENSP00000391302 [100%] APOL1-208 - ENSP00000404525 [100%]
Matching mouse transcripts	ENSMUSP00000010745 [43%] ENSMUSP00000134864 [43%] ENSMUSP00000135864 [43%] ENSMUSP00000050745 [40%] ENSMUSP00000086873 [38%]
ANTIGEN VIEWⁱ The Structure section provides in-house generated structures, predicted using the Alphafold source code, for the majority of the proteins and their related isoforms. Displaying protein features on the AlphaFold structures Individual splice variants can be selected in the top part of the Protein Browser (see below) and different transcript-related features such as transmembrane regions, InterPro domains and antigen sequences for antibodies can be displayed in the structure by clicking on the respective features in the Protein Browser. Clinical and population-based amino acid variants based on data from the Ensembl variation database and AlphaMissense (AM) predictions can be highlighted using the sliders to the right of the structure. These can also be used to colour the entire structure by residue index or make the structure autorotate.The structures are displayed using the NGL Viewer and can also be zoomed-in and rotated manually. The Protein Browser The ProteinBrowser displays the antigen location on the target protein(s) and the features of the target protein. Transcript names and schematic transcript structures including exons, introns and UTRs for the different isoforms are shown on top, and can be used to switch between the structures for the different splice variants. At the top of the view, the position of the antigen (identified by the corresponding HPA identifier) is shown as a green bar. A yellow triangle on the bar indicates a <100% sequence identity to the protein target. Below the antigens, the maximum percent sequence identity of the protein to all other proteins from other human genes is displayed, using a sliding window of 10 aa residues (HsID 10) or 50 aa residues (HsID 50). The region with the lowest possible identity is always selected for antigen design, with a maximum identity of 60% allowed for designing a single-target antigen (read more). The curve in blue displays the predicted antigenicity i.e. the tendency for different regions of the protein to generate an immune response, with peak regions being predicted to be more antigenic.The curve shows average values based on a sliding window approach using an in-house propensity scale. (read more). Signal peptides (turquoise) and membrane regions (orange) based on predictions using the majority decision methods MDM and MDSEC are also displayed. Low complexity regions are shown in yellow and InterPro regions in green. Common (purple) and unique (grey) regions between different splice variants of the gene are also displayed (read more), and at the bottom of the protein view is the protein scale.

APOL1-201 APOL1-202 APOL1-204 APOL1-205 APOL1-208

Displaying protein features on the AlphaFold structures

The Protein Browser